Cerebrospinal Fluid of Preeclamptic and Normotensive Pregnant Women Compared to Nonpregnant Women Analyzed with Mass Spectrometry

Abstract

Preeclampsia is a pregnancy-specific multiorgan disorder in which impaired placental functioning and excessive oxidative stress play an important role. We previously showed distinct differences between cerebrospinal fluid proteins in patients with preeclampsia and normotensive pregnant women. An additional group of nonpregnant women was included to study the presence of pregnancy-related proteins in normotensive and preeclamptic pregnancies and whether pregnancy-related proteins were associated with preeclampsia. Cerebrospinal fluid samples were tryptically digested and subsequently measured with a nano-LC-tribrid Orbitrap mass spectrometry system. Proteins were identified by shotgun proteomic analysis based on a data-dependent acquisition method. Proteins identified in preeclampsia, normotensive pregnant controls, and nonpregnant groups were compared to the Progenesis method according to the criteria as previously described and with a secondary analysis using a Scaffold method including Benjamini–Hochberg correction for multiple testing. For preeclampsia, the Progenesis and the Scaffold method together identified 15 (eight proteins for both analyses with one overlap) proteins that were significantly different compared to normotensive control pregnancies. Three of these 15 proteins, which were elevated in cerebrospinal fluid of preeclamptic women, were described to be pregnancy proteins with a calcium-binding function. Using two analysis methods (Progenesis and Scaffold), four out of 15 differential proteins were associated with pregnancy, as described in the literature. Three out of the four pregnancy-related proteins were elevated in preeclampsia. Furthermore, the contribution of elevated ( n = 4/15) and downregulated ( n = 2/15) calcium-binding proteins in preeclampsia is remarkably high (40%) and needs to be elucidated further.

Altered offspring neurodevelopment in an L-NAME-induced preeclampsia rat model

“…Ciampa et al demonstrated that differentially expressed proteins in the CSF of PE mothers converge to four signaling molecules, including TGF-β, vascular endothelial growth factor A, angiotensinogen, and IL-6, suggesting that neurological maternal complications of PE, such as eclampsia and posterior reversible encephalopathy syndrome, are associated with vascular remodeling, inflammation, neuronal growth, signaling, and electrophysiology ( 33 ). Güzel et al also demonstrated that some differentially expressed proteins were calcium-binding proteins, suggesting a close association between PE and calcium-binding proteins ( 34 ). As with these two studies on maternal CSF, investigation of CSF protein profiling in the offspring of the L-NAME-induced PE model may facilitate the elucidation of the mechanisms underlying subsequent neurological disorders ( 35 ).…”

Precision Localization of Lipid‐Based Nanoparticles by Dual‐Fluorescent Labeling for Accurate and High‐Resolution Imaging in Living Cells

“…ESI-LC-MS/MS: The samples (eluted in 3.5, 7, and 7.5 mL) after lipid extraction in the water phase were enzymatically digested with trypsin and subsequently measured with a nano-LC-Orbitrap MS/MS mass spectrometry system (Ultimate 3000 HPLC, Thermo Fisher Scientific Germering, Germany; Orbitrap Lumos, Thermo Fisher Scientific, San Jose, CA, USA). [61] The results were valued based on the bovine dataset. Specifically, 3 μL digest was loaded onto a C18 trap column (C18 PepMap, 300 μm inner diameter (ID) Â 5 mm, 5 μm particle size, 100 Å pore size; Thermo Fisher Scientific, The Netherlands) and desalted for 10 min using a flow rate of 20 μL min À1 0.1% TFA.…”

“…Based on this survey scan, the most intense ions were consecutively isolated (AGC target set to 10 4 ions) and fragmented by collision-activated dissociation (CAD) applying 35% normalized collision energy in the linear ion trap until a duty cycle time of 3 s was reached (top speed mode). After precursors were selected for MS/MS, they were excluded for further MS/MS spectra for 60 s. Data analysis was performed as described in the study of Güzel et al [61] Statistical Analysis: GraphPad Prism version 9 was used to perform statistical analysis. All the statistical data are depicted as mean AE standard error of the mean (S.E.M.)…”

Identification of Novel Biomarkers of Spinal Muscular Atrophy and Therapeutic Response by Proteomic and Metabolomic Profiling of Human Biological Fluid Samples

“…CDH18 has an important role in the organization of the neural circuitry and in synapse maturation [64]. CDH18 is elevated in CSF of pregnant women [65]; even if the biological meaning of its variations is unknown, this finding suggests that CDH18 is a detectable metabolite that varies with the patient's condition.…”